Do industrial solvents help antibiotic resistance?

Chinese researchers have reported that some industrial solvents may help bacteria share an antibiotic resistance gene. This means that some solvents might be leading to a rise in certain bacterial diseases.

It has been a busy week for antibiotic resistant bacteria news. First came a new report released by the World Health Organization (WHO). The report, analyzed by Pharmaceutical Microbiology, described a "post antibiotic-era—in which common infections and minor injuries can kill—far from being an apocalyptic fantasy, is instead a very real possibility for the 21st century."

Second came an update from the campaign by U.S. Senator Sherrod Brown. Brown, as the Digital Journal reported, has proposed the Strategies to Address Antimicrobial Resistance (STAAR) Act. This is legislation aimed at combating antimicrobial resistance.

In addition to these important stories, a report published in the journal Environmental Science and Technology Letters indicates that some industrial solvents may help bacteria share an antibiotic resistance gene.

While so-called “ionic liquids” are usually touted as safer substitutes for volatile organic solvents, which can release volatile organic compounds (VOCs) in household solutions, such as furniture polishes, paint stripper, and air fresheners, one ionic liquid, 1-butyl-3-methylimidazolium hexafluorophosphate (which doesn’t release VOCs), increased by 500 times the abundance of a gene that makes bacteria resistant to the antibiotic sulfonamide. If the results are confirmed or shown to occur with other compounds, another worrying set of factors will concern scientists seeking to redress antibiotic resistance.

Posted by Tim Sandle

Lonely bacteria trigger antibiotic resistance

Bacteria that are found in lower numbers are more likely to mutate, resulting in higher rates of antibiotic resistance, new research has concluded.

Researchers have drawn a connection between the environment and the ability of bacteria to develop the resistance.

Specifically, the research team discovered that the rate at which E. coli mutates depends upon how many 'friends' it has around. It seems that more lonely organisms are more likely to mutate. Related research showed that low population of bacteria developed greater resistance to the well-known antibiotic Rifampicin, used to treat tuberculosis.

From these observations, the research team argue that the change of the mutation rate is controlled by a form of social communication known as quorum sensing -- this is the way bacteria communicate to let each other know how much of a crowd there is. Quorum sensing can occur within a single bacterial species as well as between diverse species, and can regulate a host of different processes, in essence, serving as a simple indicator of population density or the diffusion rate of the bacterium’s immediate environment.

Scientists hope to build on these observations in the fight against antibiotic resistance. The study was carried out at The University of Manchester and the results have been published in the journal Nature Communications, in a paper titled “Mutation rate plasticity in rifampicin resistance depends on Escherichia coli cell–cell interactions”.

Posted by Tim Sandle

WHO endotoxin guidance

The World Health Organization is reviewing its guidelines for endotoxin testing. To examine what the changes might mean, Tim Sandle has written an article for the GMP Review.

The reference is:

Sandle, T. (2014) WHO updates endotoxin guidance for parenteral products, GMP Review, 13 (1): 7-8

For a copy, please contact Tim Sandle

Posted by Tim Sandle

Container Closure Integrity Testing

Preservation of container closure integrity (CCI) of a parenteral drug product is critical over the shelf-life of a product. Use of CCI testing is a viable alternative to sterility testing, and, according to FDA guidance, may be more useful than sterility testing in demonstrating the potential for product contamination over the shelf-life or dating period.

This important topic is examined by Louis Brasten, Barbara Jacobs, and Alicia Brydzinski for Controlled Environments magazine.

In the article, the authors note that there are many advantages to using CCI testing in lieu of sterility testing, including conservation of samples, fast results, and sensitivity that can pinpoint a leak in a vial or syringe system. It must be noted, however, that container and closure system integrity tests cannot demonstrate a product’s initial sterility.

The article can be accessed here.

Posted by Tim Sandle

WHO updates endotoxin guidance for parenteral products

The WHO has issued a draft document for the establishment of endotoxin limits for parenteral preparations. Once approved, the text will be incorporated into the International Pharmacopoeia.

Tim Sandle has written a review article into this important topic. The reference is:

Sandle, T. (2014) WHO updates endotoxin guidance for parenteral products, GMP Review, 13 (1): 7-8

For further details, please contact Tim Sandle

Posted by Tim Sandle

Multiple sclerosis 'linked to food bug'

A food poisoning bacterium may be implicated in MS, say US researchers.

Lab tests in mice by the team from Weill Cornell Medical College revealed a toxin made by a rare strain of Clostridium perfringens caused MS-like damage in the brain.

And earlier work by the same team, published in PLoS ONE, identified the toxin-producing strain of C. perfringens in a young woman with MS.

But experts urge caution, saying more work is needed to explore the link. No-one knows the exact cause of Multiple sclerosis (MS), but it is likely that a mixture of genetic and environmental factors play a role. It's a neurological condition which affects around 100,000 people in the UK.
Bacteria

C. perfringens, found in soil and contaminated undercooked meat, comes in different strains.

Most cases of human infection occur as food poisoning - diarrhoea and stomach cramps that usually resolve within a day or so. More rarely, the bacterium can cause gas gangrene.

And a particular strain of C. perfringens, Type B, which the Weill team says it identified in a human for the first time, makes a toxin that can travel through blood to the brain.

In their lab studies on rodents the researchers found that the toxin, called epsilon, crossed the blood-brain barrier and killed myelin-producing cells - the typical damage seen in MS.

Lead investigator Jennifer Linden said the findings are important because if it can be confirmed that epsilon toxin is a trigger of MS, a vaccine or antibody against the toxin might be able to halt or prevent this debilitating disease.

She presented the group's latest findings at a meeting of the American Society for Microbiology.

Dr Susan Kohlhaas, Head of Biomedical Research at the MS Society, said: "Discovering potential causes or triggers for MS could enable us to develop better treatments or even, one day, prevent the condition. This is interesting research but the findings now need to be validated in larger studies to establish if this toxin really is a potential trigger for MS."

Source: BBC Science

New name for Irish Medicines Board

The following content has just been added to The Irish Medicines Board's website

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New-Name-for-the-Irish-Medicines-Board-from-1-July-2014

On 1 July 2014, the Irish Medicines Board (IMB) will change its name to the Health Products Regulatory Authority (HPRA). We wish to give all our stakeholders advance notice so that you are aware of this change when you visit our website from 1 July and start to see documents and other materials displaying our new name and logo from that date forward. It may also be necessary for some organisations to update internal systems and records. Established in 1996, the IMB name has served us well. However, over the last 18 years our regulatory remit has expanded to include other health products...

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Posted by Tim Sandle

Order of incubation for recovery of bacteria and fungi

One of the dilemmas for the environmental monitoring of controlled environments, when a single culture medium is used, is with the order and length of incubation. The two most common regimes adopted are either 20-25^oC followed by 30-35^oC and 30-35^oC followed by 20-25^oC. There have been very few published studies to help the microbiologist with the selection of the optimal incubation regime.

Tim Sandle has written a peer-reviewed research article for the International Journal of PharmaceuticalCompounding. In the article, Dr. Sandle presents experimental data which examines the optimal order of incubation and compares recovery rates for bacteria and fungi.

The conclusion of the study is that the incubation regime has an impact on the numbers of microorganisms recovered and the incidence of recovery. Specifically, where the recovery o fungi is important, the research indicates that the 20-25^oC followed by 30-35^oC is superior.

The reference for the paper is:

Sandle, T. (2014) Examination of the Order of Incubation for the Recovery of Bacteria and Fungi from Pharmaceutical Cleanrooms, International Journal of Pharmaceutical Compounding, 18 (3): 242 – 247

Posted by Tim Sandle

Weight loss: Is the secret in your bacteria?

The search for the most successful way to shed the pounds seems never-ending.

Now researchers in China have looked at what impact gut bacteria have on people's weight.

They think that changing the type of bacteria found in the gut may be more effective at helping people to shed weight than cutting calories alone.

But can it really be as simple as that?

Experts warn we shouldn't ditch the cucumber sticks and hot yoga just yet.

The latest World Health Organization figures show there were more than 1.4 billion adults aged 20 or older who were overweight in 2008.

Of these, 200 million men and nearly 300 million women were obese.

Continue reading the main story

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Those numbers are rising - they have doubled since 1980.Lab tests in mice found an association between bacteria and obesity, but trials with people are only just getting started.

In a clinical trial, published in the journal Microbiology Ecology, scientists in Shanghai studied 93 obese people who started with an average Body Mass Index (BMI) of 32.

They fed the volunteers supplements that promoted the growth of certain types of bacteria and reduced the levels of other bacteria in the gut, alongside a balanced diet.

At 30-day, nine-week and 23-week intervals, participants filled in a questionnaire about what they had eaten in the last 24 hours and were physically examined after overnight fasting. They were also weighed and measured.

People in the trial lost on average 5kg over nine weeks, and the 45% who carried on for 23 weeks lost on average 6kg. Their average BMI fell to 29.3.

One morbidly obese patient who was studied as a precursor to the trial lost 51kg, in six months - or about a stone every three weeks.

The paper says patients' levels of the C-reactive protein, linked to clogged arteries and damage to blood vessels in the heart, was also reduced during the trial.

But the study notes: "Admittedly, the self-controlled nature of this study does not allow to infer that all described effects are due to manipulation of gut microbiota."

The diet included prebiotics, such as artichokes

No cravings?

Prof Liping Zhao, at Shanghai Jiao Tong University, worked on the study.

He says higher levels of toxin-producing bacteria, such as enterobacteria, in the gut can lead to insulin resistance, which means people would not feel full after eating one bowl of rice, instead needing five, 10 or 20 bowls to feel full.

"Their bodies were not telling them they are satisfied," he says. Prof Zhao says changing the bacteria could switch on a gene which makes the body burn fat.

He says if a person is fed a calorie-restricted diet alone, their ability to lose weight could be impaired if their gut still contains high levels of the type of bacteria, which deactivates the gene that, he says, switches on fat-burning.

He adds: "It is about time to make the public see the kind of scientific evidence we have that really shows we can be confident that gut bacteria have a pivotal role in obesity."

Bacteria changing behaviour?

But there are doubts about the drivers of the effect.

When looking at the ever-complex world of the bugs living inside us, and the nuanced way these bacteria interact with our bodies, there are concerns about adopting hard and fast rules about what does and does not work.

Dr David Weinkove, at Durham University, said the study was interesting.But he said the study did not show whether or not changing the bacteria in the diet actually caused the weight loss, or was just linked to it.

Dr Weinkove adds: "What is interesting is whether the microbes can change your behaviour, and that is causing obesity. That is a really fascinating area."

Prof Sir Stephen Bloom, at Imperial College London, says: "There are 10 times more bacteria in the body than cells. We are basically trolleys carrying bacteria around so they can be fed."

He says there is "no question" that changing the bacteria in people's bodies can have significant effects. For example, he says, dysentery introduces different bacteria into the gut and causes weight loss.

'Eat less'

Prof Bloom says it is very hard to "disentangle" what makes people lose weight when they are put on diets, as just changing what people eat tends to make them lose weight, as they are doing something different from usual.

He says people in China have different bacteria to those in the UK, so it is difficult to compare.

His advice is simple: "Eat less and take more exercise. If you eat less, you will always lose weight."

Dr Alison Tedstone, director of diet and obesity at Public Health England, says: "Overall, obesity is about eating more calories than we expend. There are many things that impact upon our gut bacteria, the key one being what we eat.

"While interesting associations are described in the study, there remain issues with the translation of the studies into meaningful advice for the general public."

She advises people to eat a "healthy, balanced diet", and be active to get to a healthy weight.

Source: BBC News

Sheep dog patrols may curb seaside bacterial infections

Border Collies may be an effective weapon against E. coli infections at the seaside according to a new study.

Researchers found that the hard working sheep dogs were successful at keeping seagulls away from beaches.

Gull dropping are known to be a source of E. coli bacteria, which can lead to abdominal cramping and diarrhoea in humans.

High levels of the bug are a leading cause of beach closures in many parts of the world, including the UK.

The bacteria are commonly found in human and animal faeces and can end up in the seas through rain water run off or from sewage.

Seagulls have been implicated in the spread of resistant versions of the bug. A recent report showed that around a third of E. coli samples taken from the birds were resistant to more than one antibiotic.

Shore patrol

In this new study, researchers assigned the dogs to 200-metre stretches of beach along the shores of Lake Michigan in the US, which were patrolled for parts of the summer season. Half way through the dogs were switched to untreated sections.

Populations of Ring-Billed gulls have soared in the region since the 1970s with numbers increasing by 10% per year.

The collies, known for their intelligence and their herding abilities, disturbed the seagulls and kept them from landing on the beaches.

"Most of the time, the dogs were kept on their leads," said Dr Elizabeth Alm from Central Michigan University, who led the study.

"They were released with the leads dropped, only when their handler directed them to chase gulls. Then the dogs were called, they would circle back, and the handler would pick the lead back up."

Over the course of the summers of 2012 and 2013, the scientists recorded the number of birds at each section of beach while water and sand samples were collected and tested for E. coli.

They found that the bacterial counts were significantly lower on those sandy stretches where the dogs had kept the gulls at bay.

Border Collies are known for their intelligence and are prized by sheep farmers for their ability to work hard

However the benefit didn't last through the whole season and the researchers found that later in the summer, bacterial numbers had risen once again. Dr Alm believes that the timing of the dog patrols is crucial to their effectiveness.

"If the E. coli establish in the sand early in the season, they appear to be able to persist, and probably even grow in the sand so that even though the dogs can remove the gulls from the beach later in the season, this late reduction in gulls does not translate in to a late season reduction in E. coli."

One key question though was the worry that the dogs themselves might increase the levels of E. coli if they had to answer nature's call while working on the beach.

"These were professional working dogs," said Dr Alm.

"They were given ample opportunity to take care of their "business" before going to work. They didn't often poop on the beach, but if they did it was immediately picked up by their handler and disposed of off the beach."

The research has been published at the annual meeting of the American Society for Microbiology.

Source: BBC Science

Book review: Pharmaceutical Regulatory Inspections

Pharmaceutical Regulatory Inspections  - Reviewed By Victor Grayson, Sterile Operations Manager, BPL (Pharmig News Number 60, p12)

Understanding the international regulatory field is an area of growing complexity and pharmaceutical facilities frequently face inspections from different global inspectorates, as well as their own national bodies. Keeping up with current Good Manufacturing Practices (GMP) can often be challenging.

To help with this complex task a new book has been produced “Pharmaceutical Regulatory Inspections”, edited by Madhu Raju Saghee. The book is timely, relevant and important.

The book contains chapters written by a number of international experts and covers everything anyone could possibly wish to know about the inspection process and quality issues affecting pharmaceuticals and healthcare.

In order to establish a baseline, the book opens with a very useful chapter by Tim Sandle and Madhu Raju Saghee which explores basic concepts of GMP and address global differences. The book then moves onto cover some key and specific topic areas including the FDA approach to GMP inspections; the current paradigm of ‘system based’ inspections and pre-approval inspections.

The book not only explains the inspection process, it also covers the key points that are likely to arise during an inspection or audit. This includes documentation, electronic systems, staff training, hygiene, licencing, compliance and so forth. The quality of the explanations is high and the book is packed full of useful examples and case studies.

Among the latter chapters of high quality is one by Siegfried Schmitt and Nabila Nazir which looks at the European inspection approach, breaking down the complexity of European regulations into a digestible format. Another useful chapter is where Tim Sandle, Madhu Raju Saghee and David Barr outline a strategy whereby organisations can respond to inspection observations in a timely, sensible and risk centric way.

For this reviewer the two gems of the book are the two closing chapters. One by Siegfried Schmitt and Richard Einig provides a survival guide for surviving inspections of API facilities; and another by Tim Sandle provides the most comprehensive guide for dealing with inspections of sterile facilities that this reviewer has ever come across.

In summary, this is an important book and it is an unique book. It is an essential item for the book shelf of anyone involved with quality inspections, audits or GMP.

Pharmaceutical Regulatory Inspections can be obtained from the publisher, Euromed

FDA Tackles Broken Generic Drug Application Process

“Today nearly 84 percent of all prescriptions filled in the U.S. are generic. The generic industry’s trade group, the Generic Pharmaceutical Association (GPhA), has released figures that show the impact of generic drugs: over the last 10 years, generic drugs have saved consumers in excess of 1.2 trillion dollars. At the same time, the FDA has been making steady progress in narrowing the gap between brand drugs and generic drugs in terms of quality and efficacy.”

This is the interesting opening part of an article by Bikash Chatterjee for Controlled Environments. The full article can be accessedhere.

Posted by Tim Sandle

Mechanism of action of amphotericin

Researchers now understand the mechanism of action of amphotericin, an antifungal drug that has been in use for more than 50 years -- even though it is nearly as toxic to human cells as it is to the microbes it attacks.

Studies have found evidence that amphotericin opens up ion channels in membranes, perhaps making them leakier to charged atoms that could disrupt a cell. Most scientists assumed that this was the drug's main mode of action. But the evidence also suggested that amphotericin interacted with sterols, such as cholesterol in animal cells and ergosterol in yeast. Rienstra and Burke focused on amphotericin's influence on sterols, hypothesizing that this might be a key to its toxicity.

The initial data supported this idea, indicating that very little of the drug -- less than 5 percent -- actually formed channels in membranes. Using nuclear magnetic resonance and other experimental tools, a research team have found that most of the amphotericin aggregates on the exterior of membranes, extracting sterols out of membranes like a sponge. Cell death follows soon after.

The research was carried out at Chad Rienstra. A report of the new findings has been published in Nature Chemical Biology:

Thomas M Anderson, Mary C Clay, Alexander G Cioffi, Katrina A Diaz, Grant S Hisao, Marcus D Tuttle, Andrew J Nieuwkoop, Gemma Comellas, Nashrah Maryum, Shu Wang, Brice E Uno, Erin L Wildeman, Tamir Gonen, Chad M Rienstra, Martin D Burke. Amphotericin forms an extramembranous and fungicidal sterol sponge. Nature Chemical Biology, 2014; DOI: 10.1038/nchembio.1496

Posted by Tim Sandle

Monitoring cleanrooms using a rapid micro method

A new paper of interest has been published.

In the paper, the researchers assessed the different operational states within a biopharmaceutical grade clean room, using a rapid microbiological method. The method was a novel system, based on spectrometry, designed for sampling, discriminating, and enumerating airborne particles (a BioVigilant IMD-A® System).

Central to the study was the aim to determine the microbiological levels as a clean room went from standard use through maintenance and shutdown, disinfection, and then back to standard use. The objective was to evaluate whether a rapid method could replace conventional environmental monitoring using growth-based media.

The paper concludes that the optical instrument can be used to process sufficient information, so that clean rooms can be returned to use following a period of unexpected downtime or following maintenance without the need to wait for the results from growth-based methods. As such, this type of rapid microbiological method is worth exploring further for clean room air monitoring.

The reference is:

Sandle, T., Leavy, C., Jindal, H. and Rhodes, R. (2014) Application of rapid microbiological methods for the risk assessment of controlled biopharmaceutical environments, Journal of Applied Microbiology, 116 (6): 1495-1505

Posted by Tim Sandle

Directory of Community Blood Centers and Hospital Blood Banks

Directory of Community Blood Centers and Hospital Blood Banks, 5th edition contains profiles of facilities from around the world. Details on volume and types of components collected, outdate rates, computer systems, governance, contact personnel and many more aspects of operations at each participating facility make this an invaluable resource.

Those involved in blood baking can compare their facility's performance with that of other institutions, network with colleagues about infrequently encountered procedures, locate where special services are offered, and find other key information to help their institution run efficiently. In addition to the facility descriptions, this volume contains listings of immunohematology reference laboratories and marrow donor centers, information on using the National Blood Exchange, and a handy chart that cross-references special services.

For further details see: Blood Bank

Posted by Tim Sandle